Human Mitochondrial DNA Damage Quantification qPCR Assay Kit

Product Description

• Detects low levels of mtDNA lesions
• Quantitative amplification of genomic DNA fragments
• Suitable for various sample types

The maintenance of mitochondrial genomic integrity is a prerequisite for proper mitochondrial function. Excessive oxidative stress can cause mitochondrial DNA (mtDNA) to be damaged by loss of its supercoiled structure. Accumulation of lesions in mtDNA is believed to be one of the causes of energy crisis in aging tissues as well as with numerous human diseases including neurodegenerative disorders, cardiovascular diseases and cancer. Damage to mtDNA is also a meaningful biomarker for evaluating genotoxicity of drugs and environmental toxins.

The RayBio® Human Mitochondrial DNA Damage Quantification Kit (MTH-DQ) is a Taqman™ probe-based qPCR assay for the specific quantitative measurement of human mitochondrial DNA damage. This kit targets the long mtDNA region (D-Loop gene), which is the most susceptible to damage. In principle, more mtDNA damage or lesions can slow down or block the progression of DNA polymerase, but intact mtDNA does not. Gene-specific Taqman™ probe qPCR is highly sensitive because of the use of “long” PCR methodology that permits the quantitative amplification of fragments of genomic DNA. As a result, very low levels of lesions can be detected, permitting the study of mtDNA damage using genomic DNA from cultured cells, tissue, saliva, scraped cells, urine, spinal fluid, lavage fluid, blood, etc.

Specificity 
The non-human and no DNA samples have higher Cq values than human DNA or are undetermined, which means there is high specificity for mtDNA amplification.

Reproducibility
CV <5% for Cq value and CV <15% for mtDNA damage when the input DNA >0.3 ng/reaction.

Performance Data

Fig.1. H₂O₂-induced mtDNA lesions. Quantification of mtDNA damage (%) by PCR amplification of total DNA isolated from Hela cells exposed to 0 – 1000 µM hydrogen peroxide (H₂O₂) for 60 minutes showing a steadily increasing mtDNA damage over H2O2 concentration in all tested mtDNA regions. Error bars designate standard deviation (four independent experiments). The input DNA amount is 1 ng/reaction for all samples.

Fig.2. Standard curves of FAM and JUN amplification for different DNA concentrations (From 30 pg - 100,000 pg/reaction) of Hela cells detected by this kit.